PROJECT SUMMARY/ABSTRACT This proposal requests funds to purchase a cutting edge Wako CV7000 High-Content Confocal Imaging system to be shared by eleven NIH-funded Principal Investigators at Stanford University School of Medicine. The new system has two purposes: it replaces a leased automated epifluorescence imaging system ImageXpress micro XL that is heavily used and it adds a critical new capability to the Stanford campus to do automated confocal imaging. The laboratories of Tobias Meyer, James Ferrell, Kang Shen, Thomas Wandless, Steven Artandi, Marius Wernig, Karlene Cimprich, Thomas Quertermous, Michael Lin, Rajat Rohatgi, and Matthew Scott are located in close proximity and share a common interest in microscopy-based dynamic analysis of cellular systems to uncover and understand the cell cycle, cell differentiation, cell damage, cell migration and other dynamic cellular processes. Central to the research in our laboratories is the use of long-term, live-cell, fluorescence microscopy to perturb, monitor and quantify dynamic changes in cellular processes. Specifically, we use methods that require automated tracking of cells, monitoring of changes in cell shape and cell polarization, of organelle distribution and of chromatin remodeling and of changes in local signaling. As part of our projects, we also need to visualize and quantify induced expression and degradation of regulatory and marker proteins as well as the movement of signaling proteins and biosensors to and from the nucleus, plasma membrane and other cellular compartments. Last year, when the manufacturer discontinued all service support for our two heavily-used, 12- year old automated ImageXpress imaging systems, we were able to obtain a one-time lease on a newer-model ImageXpress Micro XL until August 2013. This loaner system is heavily used every day 24/7 since many of our experiments take 1-3 days of continuous multi-well imaging. The advanced high-throughput, live-cell imaging capabilities of the Wako CV7000 will replace this loaned ImageXpress Micro XL and, with the critical addition of confocal capabilities, also significantly enhance the NIH-supported research in our laboratories by providing a shared resource to address our increasing microscopy needs.